Maternal prenatal depression is associated with decreased placental expression of the imprinted gene PEG3.

BACKGROUND: Maternal prenatal stress during pregnancy is associated with fetal growth restriction and adverse neurodevelopmental outcomes, which may be mediated by impaired placental function. Imprinted genes control fetal growth, placental development, adult behaviour (including maternal behaviour) and placental lactogen production. This study examined whether maternal prenatal depression was associated with aberrant placental expression of the imprinted genes paternally expressed gene 3 (PEG3), paternally expressed gene 10 (PEG10), pleckstrin homology-like domain family a member 2 (PHLDA2) and cyclin-dependent kinase inhibitor 1C (CDKN1C), and resulting impaired placental human placental lactogen (hPL) expression. METHOD: A diagnosis of depression during pregnancy was recorded from Manchester cohort participants' medical notes (n = 75). Queen Charlotte's (n = 40) and My Baby and Me study (MBAM) (n = 81) cohort participants completed the Edinburgh Postnatal Depression Scale self-rating psychometric questionnaire. Villous trophoblast tissue samples were analysed for gene expression. RESULTS: In a pilot study, diagnosed depression during pregnancy was associated with a significant reduction in placental PEG3 expression (41%, p = 0.02). In two further independent cohorts, the Queen Charlotte's and MBAM cohorts, placental PEG3 expression was also inversely associated with maternal depression scores, an association that was significant in male but not female placentas. Finally, hPL expression was significantly decreased in women with clinically diagnosed depression (44%, p < 0.05) and in those with high depression scores (31% and 21%, respectively). CONCLUSIONS: This study provides the first evidence that maternal prenatal depression is associated with changes in the placental expression of PEG3, co-incident with decreased expression of hPL. This aberrant placental gene expression could provide a possible mechanistic explanation for the co-occurrence of maternal depression, fetal growth restriction, impaired maternal behaviour and poorer offspring outcomes.

Paradoxical reactions during treatment of tuberculosis with extrapulmonary manifestations in HIV-negative patients.

PURPOSE: Tuberculous paradoxical reactions (PR) have been seldom studied in non-immunocompromised patients. We conducted a study to describe the incidence, clinical and biological features, treatment and outcome of PR in human immunodeficiency virus (HIV)-negative patients treated for extrapulmonary tuberculosis (TB) and to identify predictive factors of PR. METHODS: A single-center retrospective study was conducted in consecutive HIV-negative patients presenting with TB with at least one extrapulmonary manifestation who were hospitalized in an internal medicine department between 2000 and 2010. RESULTS: Seventy-six patients were enrolled in the study. Lymphadenitis was the most common extrapulmonary manifestation of tuberculosis among this patient population (72 %). PR occurred in 19 (25 %) patients, mostly involving the lymph nodes (68 %) and lung (16 %), but also the pericardium, pleura, bone, muscle and brain. Median time to PR onset after initiation of anti-TB regimen was 86 days (interquartile range 36-125). Treatment of PR consisted mainly of corticosteroids (47 % of patients) and needle aspiration of PR lymph nodes (31 %). Peripheral lymph node involvement (p = 0.009), lymphopenia (p = 0.03) and anemia (p = 0.002) at presentation were associated with PR occurrence. Outcome was favorable in all patients with PR but one; the latter suffered residual paraplegia. CONCLUSIONS: Paradoxical reactions are frequent in the course of extrapulmonary TB treatment in HIV-negative patients but their outcome is excellent, except in some cases with central nervous system involvement.

[Nonbacterial thrombotic endocarditis and gastric carcinoma]. / Endocardite marastique et adénocarcinome gastrique.

We report a 74-year-old woman with acute heart failure and recurrent ischemic strokes as the presenting features of a nonbacterial thrombotic endocarditis complicating a gastric adenocarcinoma. The treatment only allowed a few months remission. Diagnosis of nonbacterial thrombotic endocarditis is rarely obtained while the patient is alive. Coagulation abnormalities due to the tumoral process are responsible of the valvular thrombotic process. Anticoagulation with heparin is recommended. Valvular surgery remains controversial.

Growth-promoting effects of diabetes and insulin on arteries. An in vivo study of rat aorta.

Proliferation of arterial smooth muscle cells is regarded as an important event in atherogenesis, which according to in vitro culture studies is influenced by diabetes and insulin. To assess whether this holds true in vivo, we studied the cellular kinetics of thoracic aorta in normal and streptozocin-induced diabetic rats with and without insulin treatment. We measured the incorporation of [3H]thymidine into intima-media, as well as its DNA content, 2 and 14 days after endothelial denudation. We found that the mitotic response of an injured artery is not modified by diabetes but is depressed by insulin treatment in nondiabetic rats, probably due to hypoglycemia. Our data in insulin-treated diabetic rats support but do not definitely settle the view that insulin is mitogenic as long as the treatment does not cause sustained hypoglycemia.

[Blood pressure control after a one year follow-up in high risk subject screened in occupational medicine]. / Contrôle de la pression artérielle après un suivi d'un an chez des sujets ia haut risque cardiovasculaire dépistés en médecine du travail.

OBJECTIVE: To identify factors predicting the success or the failure of intervention on blood pressure in a population estimated at high risks. METHODS: The program "Coeur 2001" has analysed the absolute cardiovascular risk (ACVR Framingham) in 107 371 voluntary French railways employees. In the company, were considered at high risk (HR), subjects for whom risk was > or = to the 95th percentile of the distribution of the observed ACVR by age range: ACVR > or = 4.5% before 35 years, 12% between 35 and 45 years and 19% beyond 45 years, i.e. a total of 4 190 subjects. These subjects were warned about their risk and advised to choose and consult a physician. A two-year follow up was planned. Identical data (risk factors, ACVR, type of management and therapies) were collected during the first consultation with the occupational physician (T0), one year later (T1) and two years later (T2). RESULTS: Our work concerned 2376 employees at HR, consulting at T1. At T1, 54% of subjects were in the hight risk group (SHR) [48% when BP at T1 was < 140/90 mmHg and 62% when the BP was > or = 140/90 mmHg]. The mean decrease of the systolic BP (SBP) was 4 mmHg in the whole sample, 7.7 mmHg in subjects with normalised ACVR, and it remained stable in the group still at HR (-0.7 mmHg). At T0, blood pressure (BP) was > or = 140/90 mmHg in 55.8% of the patients and 38.4 at T1. This high BP was associated with higher frequency of diabetes (14 vs 7%) and overweight (BMI > or =30 kg/m2; 32.8 vs 19.7%). The percentage of treated hypertensive subjects had increased from 35 to 62% but one third of uncontrolled hypertensive subjects was treated by mono therapies at T1. CONCLUSION: To keep BP under control is a difficult task in routine medicine. At T1, despite a more aggressive treatment, 38% of subjects at high risk were still hypertensive subjects.

[Workshop to teach elderly patients how to get up from the ground: pilot prospective assessment in a hospital department of internal medicine]. / Apprentissage de la technique pour se relever du sol chez des patients âgés hospitalisés. Etude prospective en médecine interne.

BACKGROUND: Falls occur frequently among the elderly and often lead to hospitalization. Even if uninjured, some of them cannot get up again, and prolonged time on the floor and failed efforts to get up can cause complications. Training methods and exercises for getting up from the floor exist but have not been evaluated. METHODS: We set up a training course to teach elderly patients to get up from the ground and assessed it over the short term by a prospective observational study of 29 patients over 3 months. Each week, a group training session took place, followed 2-9 days later by an individual assessment conducted according to a written protocol. RESULTS: Of the 29 patients (16% of those older than 65 years admitted to our internal medicine department during this period) who received the training, 24 (80%) underwent the subsequent evaluation. Training was effective: before training, only 2 of 29 assessable patients were able to get up on their own, while at the reassessment, 11 of 24 succeeded (p=0.003). Significant improvement was observed for the first two-and most difficult-steps of the maneuver: rolling over from a supine to a prone position (p=0.003), and then moving up into a quadrupedal position (getting up onto all four limbs) (p=0.006). The only variable that appeared to predict a poor result was a Mini Mental Status (MMS) score lower than 26/30. CONCLUSION: Teaching elderly patients how to get up from the floor can be accomplished in an inpatient internal medicine department and appears to be effective in the short term. Although further studies involving more patients followed for a longer period are required to confirm and assess the actual benefits, this training is safe and can be recommended.

Influence of applied stress on mitotic response of arteries to injury with a balloon catheter: quantitative study in rat thoracic aorta.

Proliferation of arterial smooth muscle cells is an important feature of atherosclerosis, and a well documented reaction to intimal injury. To assess the influence of the intensity of injury to rat thoracic aorta, we studied the effects of a soft or hard friction with a moderately or tightly inflated balloon catheter at 2, 14 and 30 d after operation. As compared with soft injury, hard injury (1) strongly enhanced the proliferating response of the aortic intima and media (median incorporation of tritiated thymidine into deoxyribonucleic acid on day 2 increase 2.8-fold, 95% confidence interval 2.3-3.3; median deoxyribonucleic acid content on day 14; 102.1 v 72.1 micrograms); (2) markedly delayed endothelial regeneration (median percentage of intimal area stained by Evans blue on day 14: 33.1 v 0.6%). On day 2, transmission and scanning electron microscopy showed that endothelial denudation was complete after a hard injury, but only partial after a soft one. However, macroscopic staining of the intima with Evans blue was complete in both instances. The extent of endothelial denudation appears to be a major determinant of the mitotic reaction of arteries to injury. In the experimental search for drugs to reduce muscular proliferation (of potential value in the prevention of restenosis after percutaneous transluminal angioplasty), endothelial injury with balloon catheters should be carefully standardised, and applied "blindly" to afford valid comparisons between treated and control groups of animals.

Heterogeneity in migration of smooth muscle cells from normal and injured rat thoracic aorta in primary culture.

OBJECTIVE: Aside from proliferation, migration of smooth muscle cells is an essential component of the arterial sclerotic reaction. The aim of this study was to define a model to study migration. METHODS: Primary cultures of smooth muscle cells were derived from normal or injured rat thoracic aorta. An image analysis system was used to track cells migrating out of the explants and measure the displacement of their centre of gravity. RESULTS: Migration speeds for smooth muscle cells randomly sampled from the normal whole media were very heterogeneous. The media were therefore separated into three vertical segments. Cells from the middle third migrated faster than those from the upper and lower thirds, regardless of whether they originated from the anterior and posterior parts of the segment (P = 0.001). Heparin (10 micrograms.ml-1) only inhibited smooth muscle cell migration from the middle segment (P < 0.001). Migration of smooth muscle cells from explants of aorta 3 and 14 d after injury was also studied using a balloon catheter. Three days after injury, cell velocity varied widely among the segments of the same media. In contrast, 14 d after injury cells from neointimal explants migrated homogeneously and at a slower rate than those obtained from normal media. CONCLUSIONS: These experiments show migratory variations among smooth muscle cells depending upon their position in the normal aorta and their state of activation after arterial injury. This variability must be taken into account when planning experiments to study smooth muscle cell migration.

Transient down-regulation of L-type Ca(2+) channel and dystrophin expression after balloon injury in rat aortic cells.

OBJECTIVE: Migration and proliferation of arterial smooth muscle cells are critical responses during restenosis after balloon angioplasty. We investigated the changes in the expression of Ca(2+) channels and dystrophin, two determinants of contraction, after balloon injury of rat aortas. METHODS: Proliferation and migration of aortic myocytes were triggered in vivo by the passage of an inflated balloon catheter in the aortas of 12-week-old male Wistar rats. We used the whole-cell patch clamp technique to investigate Ba(2+) currents (I(Ba)) through Ca(2+) channels in single cells freshly isolated from media and neointima at various times after injury (days 2, 7, 15, 30 and 45). RESULTS: No T-type Ca(2+) channel current was recorded in any cell at any time. In contrast, a dihydropyridine (DHP)-sensitive L-type I(Ba)was recorded consistently in the media of intact aorta. After aortic injury, I(Ba) decreased dramatically (at days 2 and 7) but recovered over time to reach normal amplitude on days 30 and 45. In the neointima, I(Ba) was absent on day 15 but also increased gradually over time as observed at days 30 and 45. The use of a specific antibody directed against the L-type Ca(2+) channel alpha(1C) subunit showed, both by immunostaining and by Western blotting, no expression of the Ca(2+) channel protein on day 15. Parallel immunodetection of dystrophin showed that this marker of the contractile phenotype of SMCs was also not detectable at this stage in neointimal cells. Both proteins were re-expressed at days 45 and 63. Balloon injury induces a transient down-regulation of I(Ba) in arterial cells. CONCLUSIONS: Cell dedifferentiation and proliferation in vivo abolish the expression of L-type Ca(2+) channels and dystrophin in neointimal cells. These changes may be critical in the regulation of Ca(2+) homeostasis and, thereby, contraction of the arterial SMCs during restenosis following angioplasty.

MALDI-MS and NanoSIMS imaging techniques to study cnidarian-dinoflagellate symbioses.

Cnidarian-dinoflagellate photosynthetic symbioses are fundamental to biologically diverse and productive coral reef ecosystems. The hallmark of this symbiotic relationship is the ability of dinoflagellate symbionts to supply their cnidarian host with a wide range of nutrients. Many aspects of this association nevertheless remain poorly characterized, including the exact identity of the transferred metabolic compounds, the mechanisms that control their exchange across the host-symbiont interface, and the precise subcellular fate of the translocated materials in cnidarian tissues. This lack of knowledge is mainly attributed to difficulties in investigating such metabolic interactions both in situ, i.e. on intact symbiotic associations, and at high spatial resolution. To address these issues, we illustrate the application of two in situ and high spatial resolution molecular and ion imaging techniques-matrix-assisted laser desorption ionization mass spectrometry imaging (MALDI-MSI) and the nano-scale secondary-ion mass spectrometry (NanoSIMS) ion microprobe. These imaging techniques provide important new opportunities for the detailed investigation of many aspects of cnidarian-dinoflagellate associations, including the dynamics of cellular interactions.

Galectin-3 gene (LGALS3) expression in experimental atherosclerosis and cultured smooth muscle cells.

The galectin-3 gene (LGALS3) encodes a beta-galactose binding lectin. LGALS3 expression is associated with neoplastic transformation and with differentiation of monocytes to macrophages. Factors involved in migration, proliferation, adhesion and differentiation of vascular smooth muscle cells (SMC) play a major role during atherosclerosis development. Expression of the galectin-3 gene was not detected in quiescent SMC but was activated in aortas of hypercholesterolemic rabbits, in aortas of rats after balloon injury and in cultured SMC. These results suggest that galectin-3 production is involved in the developmental process of atherogenesis.

Partial characterization of an autoantibody recognizing the secondary binding site(s) of thrombin in a patient with recurrent spontaneous arterial thrombosis.

A patient with an 18 year history of recurrent arterial thrombosis and no evidence of atherosclerosis or embolism of cardiac origin presented with a prolonged thrombin clotting time when performed with human thrombin. The bovine thrombin clotting time was only slightly prolonged. During 30 months of follow-up, the thrombin time fluctuated, but remained prolonged. The patient has been treated with an oral anticoagulant for the past 8 years, with no thrombotic recurrence. The inhibitor activity was due to the presence of polyclonal IgGs which bound to thrombin-Sepharose. The influence of IgGs purified from the patient's serum was compared to the influence of normal IgGs in several systems exploring the catalytic activity of thrombin and the binding of the enzyme to macromolecular substrates through secondary binding site(s). We found that the IgGs did not impair the catalytic activity toward small synthetic substrates, but inhibited the binding of thrombin to fibrinogen, thrombomodulin and heparin cofactor II. Such proteins are known to require a secondary binding site of thrombin to interact with the enzyme. The anti-thrombin antibody might have resulted from an abnormal generation of thrombin. This would be the consequence of the process favouring thrombosis. Alternatively, the autoantibody might have favoured thrombosis primarily, by impairing natural antithrombotic mechanisms triggered by thrombin.

Targeting platelets containing electro-encapsulated iloprost to balloon injured aorta in rats.

Drugs can be electro-encapsulated within platelets and targeted to damaged blood vessels by exploiting the platelet's natural haemostatic properties to adhere to collagen and other vessel wall constituents revealed by injury. A rat aorta balloon angioplasty model has been used to study the effect on platelet deposition of giving iloprost loaded platelets i.v. during the balloon injury. After labelling the circulating platelets with 111-Indium before balloon injury, time course studies showed maximum platelet deposition on the injured aorta occurred at about 1 h post-injury and the deposition remained stable over the next 2-3 h. When iloprost-loaded platelets were given i.v. during injury and the circulating platelet pool labelled with 111-Indium 30 min later, platelet deposition, measured at 2 h postinjury, was substantially and significantly reduced compared with control platelet treatment. Some anti-proliferative effects of iloprost-loaded platelets given i.v. during injury have also been observed. Whereas the incorporation of [3H]-thymidine into aorta intima-media DNA at 3 days post injury was 62-fold higher in balloon injured rats than in control sham operated rats, thymidine incorporation into intima/media of rats which had received iloprost loaded platelets during injury was reduced as compared with rats subjected only to the injury procedure. The reduction was only of near significance, however, but at 14 days after injury the total DNA content of the aorta intima/media of rats given iloprost loaded platelets during injury was significantly reduced. Although iloprost loaded platelets can clearly inhibit excessive platelet deposition, other encapsulated agents may have greater anti-proliferative effects.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of local urokinase on arterial occlusion of lower limbs.

Sixty nine patients were treated with local intra-arterial urokinase (37,500 U/CTA. hr-1) for recent severe ischemia of lower limbs: 27 (40%) ultimately required amputation. The difference of amputation rate between the groups with and without thrombolysis was not significant (33% v. 42%). A biological study in 6 patients showed that local arterial plasminemia occurred in only 1 patient. Local urokinase does not strongly stimulate "endogenous" thrombolysis and enhances "exogenous" thrombolysis only very inconstantly. A better adaptation of urokinase dosage or the use of an agent with higher affinity for fibrin might improve the efficiency of local thrombolytic therapy.

Antithrombin III and heparin cofactor II in patients with chronic renal failure undergoing regular hemodialysis.

Heparin enhances the inhibition rate of thrombin by both antithrombin III (AT III) and heparin cofactor II (HC II). We studied the activity of these two plasma proteins in patients with chronic renal failure (CRF) undergoing regular hemodialysis as their heparin requirements varied widely. In 77 normal blood donors, normal ranges (mean +/- 2 SD) were 82-122% for AT III and 65-145% for HC II. When compared with these controls 82 dialyzed CRF patients had a subnormal AT III activity and a significantly (p less than 0.001) lower HC II activity. To evaluate the effect of hemodialysis we compared AT III, HC II and total proteins in plasma before and after dialysis in 24 patients (12 with normal and 12 with low basal HC II activity). AT III and HC II activities significantly (p less than 0.001) increased in absolute value. When related to total plasma proteins, in order to suppress the influence of hemoconcentration induced by dialysis, AT III decreased significantly (p less than 0.01) whereas HC II increased slightly but significantly (p less than 0.01) in the 12 patients with low initial HC II activity. The decrease of AT III induced by heparin administrated during dialysis is likely to account for this relative decrease of AT III activity. A modification of the distribution of both HC II and heparin between the vascular wall and the circulating blood is evoked to explain the relative increase in HC II activity and the need for higher heparin dosage in patients with low HC II levels.

Antithrombin III Avranches, a new variant with defective serine-protease inhibition--comparison with antithrombin III Charleville.

A decreased plasma antithrombin activity in presence or in absence of heparin was discovered in a 47-year-old patient presenting with recurrent venous thromboembolism. The immunoreactive material (AT III-IR) was normal. The same biological abnormalities were found in two relatives of the patient, leading to the diagnosis of hereditary qualitative AT III deficiency. The propositus' AT III was coeluted with normal AT III from an heparin-sepharose column. An additional step of ion-exchange chromatography on a Mono Q column using a FPLC system (Pharmacia, St-Quentin en Yvelines, France) allowed the purification of a protein which was homogenous in SDS-10% polyacrylamide electrophoresis gel (PAGE). AT III purified from propositus' plasma, normal plasma and the plasma of the patient known to have an AT III variant with defective protease binding (AT III Charleville) were compared. The specific activities measured as heparin cofactor antithrombin or factor Xa inhibition in absence of heparin were decreased to half the normal value. Kinetic studies confirmed a decreased rate of thrombin inhibition for both abnormal AT III preparations. SDS-PAGE experiments performed in purified system and immunoblots obtained from plasma showed that the two variants have different behaviour: in the case of AT III Charleville thrombin induced an apparent 5 k delta increase in molecular mass, probably due to a conformational change. AT III Avranches did not form stoechiometric complexes with thrombin, but was unmodified by the protease.

A plasma clot lysis assay based on the release of fibrin degradation products: application to the diagnosis of hypofibrinolytic states.

Using a monoclonal antibody-based assay, we measured the fibrin degradation product release in the supernatant of plasma clots obtained before and after venous occlusion (VO) in 30 patients with definite or suspected vascular thrombosis (19 definite and 2 suspected deep vein thrombosis, 6 recurrent superficial thrombophlebitis, 3 arterial occlusions of lower limbs). tPA and PAI-1 concentrations were determined using ELISA assays; the post-occlusion values were corrected for haemoconcentration. The increase in tPA during VO was correlated with haemoconcentration (r = 0.74), but 3 patients had ineffective VO (less than 2% increase in proteins). The fibrinolytic response to VO was evaluated using the shortening of the time necessary for the release of 200 micrograms of fibrin degradation products per mg of fibrinogen (delta T 200). Two among the 27 patients with effective VO were bad responders with a delta T 200 less than 3 h (whereas all the others had delta T 200 greater than 10 h). These patients had respectively a deficient tPA release (delta tPA = 1 ng/ml) and an elevated PAI-1 level at rest (33 ng/ml). Several other patients were bad responders in terms of tPA release or of shortening of the euglobulin clot lysis time but they had a normal delta T 200. This plasma clot test reflects the ability of free tPA to bind to fibrin (the amount of which depends on the level of tPA and PAI-1), and may be useful in the diagnosis of a hypofibrinolytic state.

Effects of in vitro and in vivo exposure to insulin upon glucose carbon accumulation in rat aorta: different patterns of response for intima-media and adventitia.

We have studied the effect of insulin upon the accumulation of glucose carbon into the intima-media and the adventitia of rat aorta following in vitro exposure by adding insulin to the incubation medium, and in vivo exposure by producing relative hyperinsulinemia during life and subsequently incubating excised tissue in a medium containing no insulin. Hyperinsulinemia in vivo was either (1) endogenous--following 1.5-2 hr of refeeding after a 48-hr fast--or (2) exogenous--following an i.v. injection of insulin. For adventitia, a significant stimulation was found after both in vitro (+ 53%, p < 0.001) and in vivo exposure to insulin (+ 96%, p < 0.01 with endogenous insulin; + 75%, p < 0.05 with a dosage of 2 U/kg of exogenous insulin). For intima-media, the stimulation was weak and insignificant after in vitro exposure (+ 11%, p > 0.30), but became important and significant after in vivo exposure to insulin (+ 100%, p < 0.001 with endogenous insulin; + 50%, p < 0.05 with a dosage of 0.5 U/kg and + 49%, p < 0.05 with a dosage of 2 U/kg of exogenous insulin). For the in vivo exposure experiments, we found a significant linear correlation (r = 0.611, p < 0.005) between plasma insulin concentrations and glucose carbon accumulations into intima-medias of control and refed rats. These results establish the insulin sensitivity of the adventitia and show a sensitivity of intima-media only to in vivo insulin exposure. These different behaviors of intima-media in vivo and in vitro may have a hemodynamic basis.

Insulin-like growth factor-1 stimulates vascular smooth muscle cell proliferation in rat aorta in vivo.

IGF-I is a mitogen for vascular smooth muscle cells (SMC) in vitro and enhances SMC proliferation in vivo in diabetic rats. In this study, we examined the effect of IGF-I on SMC proliferation in vivo in normal rats. Recombinant human IGF-I (0.87 and 3.1 mg/kg/day), was infused via osmotic minipumps in normal rats starting 3 days before they were subjected to aortic injury with a balloon catheter. IGF-I at an infusion rate of 3.1 mg/kg/day caused a significant increase in 3H-thymidine incorporation into DNA (+53%, P < 0.01) in the rat aortic intima-media 2 days after injury and DNA content (+13%, P < 0.05) after 11 days. The elastin and collagen contents were not changed by IGF-I infusion after 11 days. Body weight increased slightly while blood glucose was not affected. At an infusion rate of 0.87 mg/kg/day, IGF-I had no significant effects. These results suggest that circulating levels of IGF-I can stimulate SMC proliferation in vivo in normal rats but only at high concentrations.